Early diagnostic value of methylation in promoter region of FAM19A4, PAX1 and miRNA124-2 in cervical lesions

doi:10.19401/j.cnki.1007-3639.2024.08.003

Abstract

Abstract:

Background and purpose: At present, there have been many reports on the study of methylation in cervical lesions, however it is still not clinically practical as a diagnostic and shunting index of cervical lesions. In this study, we intended to investigate the early diagnostic value of FAM19A4, PAX1 and miRNA124-2 methylation in promoter region in the progression of cervical lesions. Methods: A total of 129 liquid-based cytology specimens of different grades of cervical lesions diagnosed in the Third Affiliated Hospital of Zhengzhou University from Mar. 2020 to Mar. 2022. Methylation specific PCR (MSP) was used to detect the methylation changes of FAM19A4, PAX1 and miRNA124-2 genes in different cervical lesions. Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of methylation changes of the three genes in cervical lesions. This study was approved by the Ethics Committee of the Third Affiliated Hospital of Zhengzhou University (No. 2023-135-01). Results: All the specimens were collected and divided into 4 groups according to the histological results: 42 cases of no intraepithelial lesions or malignant lesions (NILM), 28 cases of low grade squamous intraepithelial lesion (LSIL), 36 cases of high grade squamous intraepithelial lesion (HSIL), 23 cases of squamous cervical cancer (SCC). With the increase in the level of cervical lesions, FAM19A4, PAX1 and miRNA124-2 gene methylation detection rates increased gradually, and the differences were statistically significant (P<0.05). The methylation detection rates of FAM19A4, PAX1 and miRNA124-2 in HSIL group were 81.2%, 80.5% and 71.8%, respectively, and the methylation detection rates of three genes in SCC group were up to 100.0%. Cytological diagnosis of cervical cancer showed the area under curve (AUC) was 0.731, with a sensitivity and a specificity of 65.9% and 80.4%, respectively. When FAM19A4, PAX1 and miRNA124-2 were used alone to diagnose HSIL+ (HSIL and SCC), PAX1 methylation had the highest diagnostic efficiency, and the AUC was 0.925, with a sensitivity and a specificity of 92.8% and 87.3%. When the diagnosis was made in pairs, the AUC of FAM19A4 combined with PAX1 in the diagnosis of HSIL+ was 0.930, with a sensitivity and a specificity of 95.7% and 87.1%, respectively. The AUC of FAM19A4 combined with miRNA124-2 in the diagnosis of HSIL+ was 0.895, with a sensitivity and a specificity of 97.6% and 85.7%, respectively. The AUC of PAX1 combined with miRNA124-2 in the diagnosis of HSIL+ was 0.928, with a sensitivity and a specificity of 95.7% and 89.1%, respectively. When the PAX1 and FAM19A4 combined with miRNA124-2 in the diagnosis of HSIL+, the AUC was 0.928, with a sensitivity and a specificity of 100.0% and 81.8%, respectively. Conclusion: The methylation of the promoter regions of the FAM19A4, PAX1 and miRNA124-2 genes demonstrates high sensitivity and specificity in diagnosing cervical lesions, indicating the potential for becoming novel indicators for early diagnosis of cervical lesions.

Key words: FAM19A4, PAX1, miRNA124-2, Methylation, Cervical lesion

CLC Number:

R737.33

LI Ya, LIU Hui, REN Jingjing, LI Xiaofu, ZHI Yanfang. Early diagnostic value of methylation in promoter region of FAM19A4, PAX1 and miRNA124-2 in cervical lesions[J]. China Oncology, 2024, 34(8): 734-744.

Figures/Tables 14

Tab. 1

Tab. 2

Tab. 3

Tab. 4

Tab. 5

Tab. 6

Fig. 1

Tab. 7

Fig. 2

Tab. 8

Fig. 3

Tab. 9

Tab. 10

Fig. 4

References 22

[1]	TORRES-ROMAN J S, RONCEROS-CARDENAS L, VALCARCEL B, et al. Cervical cancer mortality among young women in Latin America and the Caribbean: trend analysis from 1997 to 2030[J]. BMC Public Health, 2022, 22(1): 113.
[2]	BUSKWOFIE A, DAVID-WEST G, CLARE C A. A review of cervical cancer: incidence and disparities[J]. J Natl Med Assoc, 2020, 112(2): 229-232. doi: S0027-9684(20)30043-2 pmid: 32278478
[3]	LI B H, ZHANG L, ZHAO J G, et al. The value of cytokine levels in triage and risk prediction for women with persistent high-risk human papilloma virus infection of the cervix[J]. Infect Agent Cancer, 2019, 14: 16.
[4]	EL ALIANI A, EL-ABID H, EL MALLALI Y, et al. Association between gene promoter methylation and cervical cancer development: global distribution and a meta-analysis[J]. Cancer Epidemiol Biomarkers Prev, 2021, 30(3): 450-459.
[5]	LIANG L A, EINZMANN T, FRANZEN A, et al. Cervical cancer screening: comparison of conventional pap smear test, liquid-based cytology, and human papillomavirus testing as stand-alone or cotesting strategies[J]. Cancer Epidemiol Biomarkers Prev, 2021, 30(3): 474-484.
[6]	JI W T, LOU W H, HONG Z B, et al. Genomic amplification of HPV, h-TERC and c-MYC in liquid-based cytological specimens for screening of cervical intraepithelial neoplasia and cancer[J]. Oncol Lett, 2019, 17(2): 2099-2106. doi: 10.3892/ol.2018.9825 pmid: 30675277
[7]	VILLANUEVA L, ÁLVAREZ-ERRICO D, ESTELLER M. The contribution of epigenetics to cancer immunotherapy[J]. Trends Immunol, 2020, 41(8): 676-691. doi: S1471-4906(20)30126-5 pmid: 32622854
[8]	HAINES K, XU F, VAN ARSDALE A, et al. Investigation of perturbation of DNMT and TET enzymes in endometrial cancer (308)[J]. Gynecol Oncol, 2022, 166: S161-S162.
[9]	KELLY H, BENAVENTE Y, PAVON M A, et al. Performance of DNA methylation assays for detection of high-grade cervical intraepithelial neoplasia (CIN2+): a systematic review and meta-analysis[J]. Br J Cancer, 2019, 121(11): 954-965.
[10]	FANG C, WANG S Y, LIOU Y L, et al. The promising role of PAX1 (aliases: HUP48, OFC2) gene methylation in cancer screening[J]. Mol Genet Genomic Med, 2019, 7(3): e506.
[11]	CHENG S J, CHANG C F, KO H H, et al. Hypermethylated ZNF582 and PAX1 genes in oral scrapings collected from cancer-adjacent normal oral mucosal sites are associated with aggressive progression and poor prognosis of oral cancer[J]. Oral Oncol, 2017, 75: 169-177.
[12]	SMOLARZ B, DURCZYŃSKI A, ROMANOWICZ H, et al. The role of microRNA in pancreatic cancer[J]. Biomedicines, 2021, 9(10): 1322.
[13]	The clinical application value of FAM19A4/mir124-2 methy lation test in hrHPV-positive women[J]. Cancer Cell Res, 2022, 9(33).
[14]	KUMAR R, PAUL A M, RAMESHWAR P, et al. Epigenetic dysregulation at the crossroad of women’s cancer[J]. Cancers, 2019, 11(8): 1193.
[15]	XU W X, XU M Y, WANG L L, et al. Integrative analysis of DNA methylation and gene expression identified cervical cancer-specific diagnostic biomarkers[J]. Signal Transduct Target Ther, 2019, 4: 55.
[16]	GUO M Z, PENG Y J, GAO A A, et al. Epigenetic heterogeneity in cancer[J]. Biomark Res, 2019, 7: 23. doi: 10.1186/s40364-019-0174-y pmid: 31695915
[17]	LI N, HU Y J, ZHANG X Y, et al. DNA methylation markers as triage test for the early identification of cervical lesions in a Chinese population[J]. Int J Cancer, 2021, 148(7): 1768-1777. doi: 10.1002/ijc.33430 pmid: 33300604
[18]	DVORSKÁ D, BRANÝ D, NAGY B, et al. Aberrant methylation status of tumour suppressor genes in ovarian cancer tissue and paired plasma samples[J]. Int J Mol Sci, 2019, 20(17): 4119.
[19]	周俏苗, 汪洪林, 黄海燕, 等. MiR-124-3p靶向调控MAPK 14对子痫前期大鼠胎盘滋养层细胞增殖及侵袭的影响[J]. 中国比较医学杂志, 2020, 30(6): 1-9.
	ZHOU Q M, WANG H L, HUANG H Y, et al. MiR-124-3p affects proliferation and migration in preeclampsia pathogenesis by targeting MAPK 14[J]. Chin J Comp Med, 2020, 30(6): 1-9.
[20]	JIA X Q, WANG X, GUO X R, et al. MicroRNA-124: an emerging therapeutic target in cancer[J]. Cancer Med, 2019, 8(12): 5638-5650.
[21]	SHIMIZU T, SUZUKI H, NOJIMA M, et al. Methylation of a panel of microRNA genes is a novel biomarker for detection of bladder cancer[J]. Eur Urol, 2013, 63(6): 1091-1100. doi: 10.1016/j.eururo.2012.11.030 pmid: 23200812
[22]	ADCOCK R, NEDJAI B, LORINCZ A T, et al. DNA methylation testing with S5 for triage of high-risk HPV positive women[J]. Int J Cancer, 2022, 151(7): 993-1004. doi: 10.1002/ijc.34050 pmid: 35477862

Group	Pathological diagnosis				Total	x² value	P value
Group	NILM	LSIL	HSIL	SCC	Total	x² value	P value
Age/year	37.00±11.58	40.90±12.27	42.80±9.84	53.40±12.69^*	42.22±11.59	8.901	0.000
HPV
Positive	47.6% (20/42)	89.3% (25/28)	97.2% (35/36)	91.3% (21/23)	78.3% (101/129)	35.126	0.000
Negative	52.4% (22/42)	10.7% (3/28)	2.8% (1/36)	8.7% (2/23)	21.7% (28/129)
TCT
NILM	42				42
ASC-US		9	3		12
ASC-H			2	3	5
LSIL		19	2		21
HSIL			29	11	40
SCC				9	9

Gene	NILM (n=42)	LSIL (n=28)	HSIL (n=36)	SCC (n=23)	χ² value
PAX1	21.4% (9/42)	35.7% (10/28)	80.5% (29/36)	100.0% (23/23)	51.659
miRNA124-2	28.5% (12/42)	29.6% (8/27)	71.8% (23/32)	100.0% (23/23)	40.975
FAM19A4	45.2% (19/42)	51.8% (14/27)	81.2% (26/32)	100.0% (23/23)	25.688

Gene	NILM (n=42)	LSIL (n=27)	HSIL (n=32)	SCC (n=23)	χ² value
P+miR	38.1% (16/42)	44.4% (12/27)	84.3% (27/32)	100% (23/23)	34.906
P+F	47.6% (20/42)	59.3% (16/27)	87.5% (28/32)	100% (23/23)	26.104
F+miR	54.7% (23/42)	59.3% (16/27)	81.2% (26/32)	100% (23/23)	18.201
P+F+miR	57.1% (24/42)	66.7% (18/27)	87.5% (28/32)	100% (23/23)	18.476

Item	FAM19A4	PAX1	miR124-2	P+F	P+miR	miR+F	P+F+miR
Sensitivity	81.25%	80.56%	71.88%	87.50%	84.38%	81.25%	87.50%
Specificity	81.25%	78.57%	71.43%	52.38%	61.90%	45.24%	42.86%
Total coincidence rate	66.22%	79.49%	71.62%	67.57%	71.62%	60.81%	62.16%

Item	PAX1	FAM19A4	miR124-2	P+F	P+miR	miR+F	P+F+miR
Sensitivity	88.14%	89.09%	83.64%	92.73%	90.91%	89.09%	92.73%
Specificity	72.86%	52.17%	71.01%	47.83%	59.42%	43.48%	39.13%
Total coincidence rate	79.84%	68.55%	76.61%	67.74%	73.39%	63.71%	62.90%